Analysis of mouse glomerular podocyte mRNA by single-cell reverse transcription-polymerase chain reaction.
Selective investigation of glomerular podocytes is not possible using conventional methods in vivo. Analysis of glomerular epithelium-derived cells in culture yields dubious results because of the rapid dedifferentiation of podocytes. We developed a modification of the polymerase chain reaction (PCR) method previously used to analyze cultured neurons. Podocytes harvested from freshly dissected glomeruli are ideal target cells for this modified, single cell reverse transcription-PCR method to reproducibly identify specific mRNA species from resident intact podocytes in vivo.
Schröppel
B
B
Medizinische Poliklinik, Ludwig-Maximilians-Universität, München, Germany.
Huber
S
S
Horster
M
M
Schlöndorff
D
D
Kretzler
M
M
D000199:Actins / Q000235:genetics; D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics; D016228:Endothelial Growth Factors / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism*; D008222:Lymphokines / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D016133:Polymerase Chain Reaction; D017027:Protein Tyrosine Phosphatases / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis*; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D025721:WT1 Proteins
D000199:Actins; D004268:DNA-Binding Proteins; D016228:Endothelial Growth Factors; D008222:Lymphokines; D008565:Membrane Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D025721:WT1 Proteins; D017027:Protein Tyrosine Phosphatases; C516890:Ptpro protein, mouse; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3